Immobilization of Candida cylindracea lipase on poly lactic acid, polyvinyl alcohol and chitosan based ternary blend film: Characterization, activity, stability and its application for N-acylation reactions

• Published in: Process biochemistry (1991) 2013-09, Vol.48 (9), p.1335-1347
• Main Authors: Badgujar, Kirtikumar C., Dhake, Kishor P., Bhanage, Bhalchandra M.
• Format: Article
• Language: English
• Subjects: Acetates; Biocatalysis; Blends; Candida cylindracea; Candida cylindracea lipase (CCL); Chitosan; Entrapment; Immobilization; Lipase; N-acylation; Orbitals; Polyvinyl alcohols; Ternary blend polymer film
• ISSN: 1359-5113; 1873-3298
• DOI: 10.1016/j.procbio.2013.06.009

•PLA, CH and PVA were used to make biocompatible ternary blend polymer film.•Ternary blend polymer film was used to immobilize Candida cylindracea lipase.•SEM, IR, DSC, % water content analysis was done to characterize immobilized lipase.•Reaction conditions were studied and applied for enzymatic synthesis of acetamide.•Immobilized CCL was well reused up to four cycles and notably stable up to 300 days. The ecofriendly ternary blend polymer film was prepared from the chitosan (CH), polylactic acid (PLA) and polyvinyl alcohol (PVA). Immobilization of Candida cylindracea lipase (CCL) was carried out on ternary blend polymer via entrapment methodology. The ternary blend polymer and immobilized biocatalyst were characterized by using N2 adsorption–desorption isotherm, SEM, FTIR, DSC, and (%) water content analysis through Karl Fischer technique. Biocatalyst was then subjected for the determination of practical immobilization yield, protein loading and specific activity. Immobilized biocatalyst was further applied for the determination of biocatalytic activity for N-acylation reactions. Various reaction parameters were studied such as effect of immobilization support (ratio of PLA:PVA:CH), molar ratio (dibutylamine:vinyl acetate), solvent, biocatalyst loading, time, temperature, and orbital speed rotation. The developed protocol was then applied for the N-acylation reactions to synthesize several industrially important acetamides with excellent yields. Interestingly, immobilized lipase showed fivefold higher catalytic activity and better thermal stability than the crude extract lipase CCL. Furthermore various kinetic and thermodynamic parameters were studied and the biocatalyst was efficiently recycled for four successive reuses. It is noteworthy to mention that immobilized biocatalyst was stable for period of 300 days.